Use of cationic preservative in food products

ABSTRACT

A novel use of cationic preservatives and preparations according to this novel use. A cationic preservative derived from lauric acid and arginine, in particular, the ethyl ester of the lauramide of the arginine monohydrochloride, hereafter named LAE, can be used for the protection against the growth of microorganisms. LAE and related compounds are particularly suitable to be used in the preservation of all perishable food products. The composition optionally comprises auxiliary components and excipients.

RELATED APPLICATIONS

This application is a continuation of, and claims priority under 35U.S.C. §120 to, and commonly-assigned U.S. application Ser. No.10/493,783 which was filed on Apr. 26, 2004 now U.S. Pat. No. 7,407,679and which is a national phase of International Application No.PCT/EP01/12358 filed on Oct. 25, 2001. The entire disclosures of theseearlier applications are hereby incorporated by reference.

This invention relates to a novel use of cationic preservatives andpreparations according to this novel use.

Despite the food industry must avoid the use of preservative products bymeans of good manufacture practices as it is described in the nationaland international regulations, it is often necessary to warrant theneeded storage capability of the produced food-stuff but never to hidedefective effects of a manipulation or manufacture technique.

A cationic preservative derived from lauric acid and arginine, inparticular, the ethyl ester of the lauramide of the argininemonohydrochloride, hereafter named LAE, could be used for the protectionagainst the growth of the microorganisms. The chemical structure isdescribed in the following formula:

The preparation of this product has been described in Spanish patentapplication ES-A-512643.

Biological studies carried out at different research centres undersupervision of the present applicant showed LAE to act mainly over theexternal and cytoplasmatic membrane of the microorganisms and, also,into the cytoplasmatic media, preventing their proliferation. Its actiondepends on the kind of microorganism and on the exposure time.

Besides, its metabolism in rats has been studied showing a fastabsorption and metabolisation into naturally-occurring amino acids andthe fatty acid lauric acid, which are eventually excreted as carbondioxide and urea. Toxicological studies have demonstrated, that LAE iscompletely harmless to animals and humans.

Therefore, LAE and related compounds are particularly suitable to beused in the preservation of all perishable food products.

This compound is remarkable for its inhibitory action over theproliferation of different microorganisms, such as bacteria, fungi andyeasts. The minimum inhibitory concentrations of LAE are shown in thefollowing table 1.

TABLE 1 M.I.C. Kind Microorganism (ppm) Gram + Arthrobacter oxydans ATCC8010 64 Bacteria Bacillus cereus var mycoide ATCC 11778 32 Bacillussubtilis ATCC 6633 16 Clostridium perfringens ATCC 77454 16 Listeriamonocytogenes ATCC 7644 10 Staphylococcus aureus ATCC 6538 32Micrococcus luteus ATCC 9631 128 Lactobacillus delbrueckii ssp lactisCECT 372 16 Leuconostoc mesenteroides CETC 912 32 Gram − Alcaligenesfaecalis ATCC 8750 64 Bacteria Bordetella bronchiseptica ATCC 4617 128Citrobacter freundii ATCC 22636 64 Enterobacter aerogenes CECT 689 32Escherichia coli ATCC 8739 32 Escherichia coli 0157H7 20 Klebsiellapneumoniae var pneumoniae CECT 178 32 Proteus mirabilis CECT 170 32Pseudomonas aeruginosa ATCC 9027 64 Salmonella typhimurium ATCC 16028 32Serratia marcenses CECT 274 32 Mycobacterium phlei ATCC 41423 2 FungiAspergillus niger ATCC14604 32 Aureobasidium pullulans ATCC 9348 16Gliocadium virens ATCC 4645 32 Chaetonium globosum ATCC 6205 16Penicillium chrysogenum CECT 2802 128 Penicillium funiculosum CECT 291416 Yeast Candida albicans ATCC 10231 16 Rhodotorula rubra CECT 1158 16Saccharomyces cerevisiae ATCC 9763 32

The use of the invention relates to cationic preservatives derived fromthe condensation of fatty acids and esterified dibasic amino acids,according to the following formula:

where:X⁻ is Br⁻, Cl⁻, or HSO₄ ⁻R₁: is a linear alkyl chain from a saturated fatty acid or hydroxyacidfrom 8 to 14 atoms of carbon bonded to the α-amino acid group through anamidic bond.R₂: is a linear or branched alkyl chain from 1 to 18 carbon atoms or anaromatic group.R₃: is:

and n can be from 0 to 4.

The most preferred compound of the above class of compounds is LAE.

It is preferred to dissolve the compound directly before use in one ofthe following preferred solvents of food grade: water, ethanol,propylene glycol, isopropyl alcohol, other glycols, mixtures of glycolsand mixtures of glycols and water. If the treatment shall be performedat a specific pH value the use of a corresponding buffer solution may berecommendable.

The composition optionally comprises auxiliary components andexcipients. Such auxiliary components and excipients can be thickeningagents (e.g. xanthan gum, guar gum, modified starches), anti-foam agents(e.g. dimethylpolysiloxane, silicon dioxide), products to obtain theoptimal pH value (e.g. phosphates, tartrates, citrates, lactates),colouring agents (e.g. curcumin, tartrazine, erythrosine), and aromaproducts. It is preferred, that the preservative composition comprisesLAE in an amount of from 0.0001% to 1% by weight relative to the wholeweight of the preservative composition.

It is particularly preferred to use the inventive composition for thepreservation of meat products, like for instance meat, poultry products,fish, crustaceans, vegetables, greens, emulsions, sauces, confectionery,bakery, pre-cooked meals, ready-to-serve meals, dairy products,egg-based products, jams, jellies, beverages, juices, wines and beers.

Moreover, the intended use relates to: wine-based flavoured drinksincluding products; non-alcoholic flavoured drinks; liquid teaconcentrates and liquid fruit and herbal infusion concentrates; BarleyWater; fruit and citric juices; Capilé Groselha; grape juices,unfermented, for sacramental use; wines, alcohol-free wines, fruit wines(including alcohol-free), alcoholic drinks with fruit; made wines, fruitsparkling wines, ciders, beers and perries (including alcohol-free);fermentation vinager; sod, salt; mead; spirits with less than 15%alcohol by volume; fillings of ravioli and similar products; quince,jams, jellies, marmalades and other fruit based spreads, candied,crystallized and glacé fruit and vegetables; sugar, glucose syrup,molasses and other sugars; transformed and dried fruits and vegetables,Frugtgrod and Rote Grütze, fruit and vegetable preparations (includingfruit-based sauces); vegetable flesh; shell fruits; mousse, compote;salads, fruits and similar products, canned or bottled; Mostarda DiFruta; Mascarpone; fruit based cake fillings; fruit gelling extracts andliquid pectine; vegetables and fruits in vinegar, brine or oil;rehydrated dried fruits; dressed dried fruits; sweetcorn canned invacuum; potato dough and pre-fried, sliced, transformed, frozen,deep-frozen and peeled potatoes; dehydrated potato flakes and granulated(?); gnocchi; polenta; olives and olive-based preparations; jellycoating of meat products (cooked, cured or dried); burger meat;heat-treated meat products, sausages, breakfast sausages, pickled porks,pates, Foie Gras, Foie Gras Entier, Blocs de Foie Gras; Sagu; Mehu andMakeutettu; Ostkaka; Pasha; Semmelknodelteig; Polsebrod and bolleryDansk; canned Flutes; gelatine; collagen based covers with a wateractivity of more than 0.6; salted meats, cured placenta, dried meatproducts; semi-preserved fish products including fish roe products,pickling, salted, dried fish, shrimps, cooked, Crangon crangon andCrangon vulgaris cooked; fresh, cooked, frozen and deep-frozencrustacean; cheese, pre-packed, sliced, unripened and cured cheese,processed cheese, layered cheese and cheese with added foodstuffs;superficial treatment of cheese, fruits and vegetables; cheesesubstitute, meat substitute, fish substitute, crustacean substitute;non-heat-treated dairy-based desserts, curdled milk, semolina andtapioca based desserts; liquid egg (white, yolk or whole egg),dehydrated, concentrated, frozen and deep-frozen egg products;pre-packed and sliced bread and rye-bread; partially baked, pre-packedbakery wares intended for retail sale, fine bakery wares with a wateractivity of more than 0.65; low-energetic bread; dry-biscuits; cereal-or potato-based snacks and coated nuts; batters, confectionery, glucosesyrup based confectionery, flour based confectionery with a wateractivity of more than 0.65, chewing gum; Christmas pudding, nougats andmarzipans; clotted cream; toppings (syrups for pancakes, flavouredsyrups for milkshakes and ice cream, similar products), fat emulsions,dressing salads, emulsified sauces, non-emulsified sauces; preparedsalads, mustard, seasonings and condiments; liquid soups and broths;aspic, liquid dietary food supplements; pearl barley; dietetic foodsintended for special medical purposes and starches; dietetic formulaefor weight control intended to replace total daily food intake or anindividual meal; and other food products where the use of preservativesbecame necessary and allowed by law.

The cationic preservative may be added to a final stage of the productto be preserved or it may be added to a initial stage which would havethe advantage of treating the food product, whereby it may be added asdry product to the product to be preserved, or in the form of a solutionor dispersion.

The food products according to the invention are prepared according tothe techniques which are well known to a person skilled in the art.

Procedures to Determine the Microbiological Population and PreservativeEffect

The determination of the microbiological population is carried outaccording to the ISO standards.

The samples are treated by dilution in buffer peptone with theappropriate neutralising agent of the preservative. The culture mediaused for counting the microorganisms are: Soya triptone agar (32-35° C.,48 hours) for the determination of mesophilic bacteria; Sabouraud agarwith chloramfenicol (25° C., 3-5 days) for fungi and yeast; Violet redbile glucose agar (32-35° C., 24 hours) for enterobacteria; Soyatriptone agar (17° C., 5 days) for psychrotrophic bacteria.

EXAMPLES

Different examples of food products and formulations are shown where theproduct has been assayed. These examples are a part of the preparationsand formulations assayed.

Example 1

This example shows the use of LAE in semi-preserved codfish in oil(table 2). The sample LAE was added to the oil assayed at aconcentration of 100 ppm and its microbiological evolution at 4° C. wascompared against a control.

TABLE 2 Time (days) 0 14 43 Microorganism Aerobe Mould & Aerobe Mould &Aerobe Mould & Bacteria yeast Bacteria yeast Bacteria yeast SampleControl (cfu/g) 3.4 · 10³ 4.0 · 10² 3.8 · 10⁵ 2.0 · 10⁴ 2.7 · 10⁸ 1.2 ·10⁷ With LAE (cfu/g) 7.6 · 10³ 3.0 · 10² 1.0 · 10⁴ 5.4 · 10³ 8.5 · 10⁴8.2 · 10³

Example 2

This example shows the use of LAE in a chicken product (table 3). Thesample LAE was added to at a concentration of 150 ppm and the evolutionof aerobe and psychrotrophic bacteria at 10° C. was compared against acontrol.

TABLE 3 Time (days) 0 14 43 Microorganism Aerobe Psychro. AerobePsychro. Aerobe Psychro. Bacteria Bacteria Bacteria Bacteria BacteriaBacteria Sample Control (cfu/g) 3.1 · 10⁵ 2.4 · 10⁴ 9.8 · 10⁵ 6.5 · 10⁵7.5 · 10⁸ 4.2 · 10⁸ With LAE (cfu/g) 1.2 · 10⁵ 3.0 · 10⁴ 4.2 · 10⁵ 7.1 ·10⁴ 6.1 · 10⁵ 6.8 · 10⁴

Example 3

This example shows the use of LAE in a carbonated orange beverage (table4). The sample LAE was added to at a concentration of 100 ppm and itsmicrobiological evolution at 17° C. was compared against a control.

TABLE 4 Time (days) 0 14 Microorganism Aerobe Mould & Aerobe Mould &Bacteria yeast Bacteria yeast Sample Control 4.0 · 10² <10 6.5 · 10⁴ 1.7· 10³ (cfu/g) With LAE 4.3 · 10² <10 1.0 · 10² <10 (cfu/g)

Example 4

This example shows the use of LAE in a blackberry juice (table 5). Thesample LAE was added to at a concentration of 60 ppm and itsmicrobiological evolution at 34° C. was compared against a control.

TABLE 5 Time (days) 0 14 Microorganism Aerobe Mould & Aerobe Mould &Bacteria yeast Bacteria yeast Sample Control 5.1 · 10² <10 2.5 · 10⁵ 3.7· 10⁴ (cfu/g) With LAE 4.0 · 10² <10 2.4 · 10³ <10 (cfu/g)

Example 5

This example shows the use of LAE in custard (table 6). The sample LAEwas added to at a concentration of 100 ppm and its microbiologicalevolution at 25° C. was compared against a control.

TABLE 6 Time (days) 0 5 Microorganism Aerobe Anaerobe Aerobe AnaerobeBacteria Bacteria Bacteria Bacteria Sample Control (cfu/g) <10 <10 9.1 ·10⁷ 3.4 · 10⁷ With LAE (cfu/g) <10 <10 1.1 · 10³ 4.1 · 10²

Example 6

This example shows the use of LAE in fairy cakes (table 7). The sampleLAE was added to at a concentration of 80 ppm and its microbiologicalevolution at 25° C. was compared against a control.

TABLE 7 Time (months) 0 3 Microorganism Aerobe Mould & Aerobe Mould &Bacteria yeast Bacteria yeast Sample Control (cfu/g) <10 <10 9.1 · 10⁴3.4 · 10³ With LAE (cfu/g) <10 <10 1.1 · 10² <10

Example 7

This example shows the use of LAE in veal stew (table 8). The sample LAEwas added to at a concentration of 100 ppm and its microbiologicalevolution at 10° C. was compared against a control.

TABLE 8 Time (days) 0 14 Microorganism Aerobe Mould & Entero- AerobeMould & Entero- Bacteria yeast bacteria Bacteria yeast bacteria SampleControl (cfu/g) <10 <10 <10 9.1 · 10⁴ 3.4 · 10³ 1.1 · 10² With LAE(cfu/g) <10 <10 <10 <10 <10 <10

Example 8

This example shows the use of LAE in ketchup (table 9). The sample LAEwas added to at a concentration of 100 ppm and its microbiologicalevolution at 25° C. was compared against a control.

TABLE 9 Time (days) 0 14 Microorganism Aerobe Mould & Entero- AerobeMould & Entero- Bacteria yeast bacteria Bacteria yeast bacteria SampleControl (cfu/g) <10 <10 <10 1.2 · 10⁶ 4.3 · 10² 1.4 · 10³ With LAE(cfu/g) <10 <10 <10 2.2 · 10³ 1.4 · 10¹ <10

The invention claimed is:
 1. A food product containing a cationicpreservative which is the ethyl ester of lauramide of argininehydrochloride having the following formula:

wherein said cationic preservative is present in the food product in anamount from about 0.0001% to about 0.006% by weight.
 2. The food productof claim 1 wherein the food product comprises fish, crustaceans, fishsubstitutes or crustacean substitutes.
 3. The food product of claim 1wherein the food product comprises meat, meat substitutes or poultryproducts.
 4. The food product of claim 1 wherein the food productcomprises vegetables, greens, sauces or emulsions.
 5. The food productof claim 1 wherein the food product comprises beverages, juices, winesor beers.
 6. The food product of claim 1 wherein the food productcomprises dairy, egg-based, jam, jelly, bakery or confectionaryproducts.
 7. The food product of claim 1 wherein the food productcomprises pre-cooked meal or ready-to-serve meal products.
 8. A methodof preservation of a food product comprising the step of adding acationic preservative which is the ethyl ester of lauramide or argininehydrochloride having the following formula:

wherein said cationic preservative is added to the food product as asolution, dispersion or solid before, during and/or after themanufacture of the food product at a concentration of from about 0.0001%to about 0.006% by weight.
 9. The method of claim 8 wherein the foodproduct comprises fish, crustaceans, fish substitutes or crustaceansubstitutes.
 10. The method of claim 8 wherein the food productcomprises meat, meat substitutes or poultry products.
 11. The method ofclaim 8 wherein the food product comprises vegetables, greens, sauces oremulsions.
 12. The method of claim 8 wherein the food product comprisesbeverages, juices, wines or beers.
 13. The method of claim 8 wherein thefood product comprises dairy, egg-based, jam, jelly, bakery orconfectionary products.
 14. The method of claim 8 wherein the foodproduct comprises pre-cooked meal or ready-to-serve meal products.
 15. Amethod of preservation of food products, comprising the step of addingto the food product a cationic preservative which is the ethyl ester oflauramide of arginine hydrochloride having the following formula:

wherein said cationic preservative is applied by surface treatmentbefore, during and/or after the manufacture of the food product at aconcentration of from about 0.0001% to about 0.006% by weight.
 16. Themethod of claim 15 wherein the cationic preservative is applied byspraying.